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The present study was aimed to evaluate the effect of seasonal variation in Temperature Humidity Index (THI) on milk production and composition in Murrah buffaloes. Eight adult lactating Murrah buffaloes of 3-5 years of age and insecond to fourth lactation were selected in summer and winter season. Meteorological variables such as ambient temperature and relative humidity were recorded and THI was calculated during the period of study. Milk samples were collected from Murrah buffaloes and evaluated for the concentration of milk fat, protein, SNF, lactose and salt. The data was analyzed using SPSS v.20. The results indicated that average milk production, milk fat, protein and SNF decreased significantly (p<0.05) in summer in comparison to winter, while lactose and salt did not differ significantly (p>0.05) in between seasons. From the present study it was concluded that high THI imposes significant heat stress and negatively affects milk production and composition in Murrah buffaloes. Hence, additional feed, shelter and management practices might be adopted to overcome the negative effects of thermal stress and also to optimize milk production of the animals.

The present study aimed to evaluate the effect of α‐tocopherol on viability, lipid peroxidation and the expression of apoptosis, stress and development‐related genes in the vitrified sheep secondary follicles. Ovarian secondary follicles (200–300 μm) were isolated and distributed separately to the vitrification treatment and supplemented with 5 mM, 10 mM, 20 mM and 30 mM of α‐tocopherol (while the control fresh group was without vitrification and supplementation of α‐tocopherol). After a week, the follicles were thawed and evaluated for follicular viability by trypan blue dye exclusion method, lipid peroxidation and gene expression studies. The results showed that the vitrification with 10 and 20 mM of α‐tocopherol positively affected (p < .05) the viability of vitrified follicles in comparison with vitrified ones without α‐tocopherol but the higher concentration of α‐tocopherol, i.e., 30 mM negatively affected the viability (p < .05) in comparison with the 10 and 20 mM of α‐tocopherol groups. The malondialdehyde (MDA) levels were significantly (p < .05) higher in the vitrified without α‐tocopherol group in comparison to the vitrified with 20 mM of α‐tocopherol group. The expression of apoptotic‐related gene, BCL2L1 was significantly higher in 10 mM α‐tocopherol group compared to the control fresh and CASPASE 3, 9 expressions were significantly higher in the vitrified group when compared to the vitrified with 10 mM α‐tocopherol group. Expressions of BAX, BAD, BAK, BMP‐15 and GDF‐9 showed no significant difference among the groups. The mRNA expression of SOD1 was significantly higher in the vitrified without α‐tocopherol group when compared to other groups. We conclude that the supplementation of 10 and 20 mM α‐tocopherol in vitrification solution was the efficient vitrification procedure for the vitrification of ovine secondary follicles.

BACKGROUND: Vitrification increases the production of reactive oxygen species (ROS) and the antioxidants in the vitrification solution may be beneficial by reducing excessive ROS production. OBJECTIVE: To evaluate the effect of retinol supplementation in vitrification solution on viability, apoptosis and development-related gene expression in vitrified sheep preantral follicles. MATERIALS AND METHODS: Preantral follicles were isolated and randomly assigned into one of five groups: Group1, control fresh preantral follicles; Group 2, vitrification treatment; Group 3, vitrification + 2 μM retinol; Group 4, vitrification + 5 μM retinol ; Group 5, vitrification + 10 μM retinol . Preantral follicles were placed in vitrification solutions and then plunged into liquid nitrogen (-196°C). After a week, the follicles were thawed and analyzed for follicular viability by trypan blue exclusion method and for gene expression. RESULTS: Vitrification with 5 μM retinol positively affected viability in comparison with vitrification without retinol (P < 0.05). There was no significant difference in viability among the Group 1, Group 2, Group 3 and Group 5. Expression of apoptotic genes BAX and Casp 3 were higher in the vitrified group, and vitrification with 5 μM retinol (Group 4) is comparable to the control fresh. Expressions of other apoptosis-related genes (i. e., BCL2L1, BAD and BAK) showed significant difference between the control fresh group and the vitrification group with 5 μM retinol. Expression of Annexin5 was also significantly different among various groups. The expression of development competence genes GDF-9 and BMP-15 were higher (P < 0.05) in the Group vitrified with 5 μM retinol. CONCLUSION: The supplementation of 5 μM retinol in vitrification solution was beneficial for the vitrification of ovine preantral follicles.

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